基于PCRarray技术探讨脾虚痰浊AS对心肌线粒体能量代谢通路相关基因mRNA表达的影响

Based on PCR Array Technology Investigating Effect of Spleen Phlegm Atherosclerosis Related mRNA Expression in Myocardial Mitochondrial Energy Metabolism Pathway

目的:利用PCRarray的技术检测脾虚痰浊动脉粥样硬化(AS)巴马小型猪心肌线粒体能量代谢信号通路相关基因mRNA的表达。方法:普通级雄性巴马小型猪10只,随机分为正常组,脾虚痰浊AS组。脾虚痰浊AS组予饮食不节、劳倦过度结合高脂饲料饮食。使用全自动生化分析仪测定TC、TG、LDL-C水平,提取各组巴马小型猪心肌总RNA,应用线粒体能量代谢信号通路PCR芯片测定脾虚痰浊AS巴马小型猪线粒体能量代谢通路相关mRNA表达。结果:各组巴马小型猪血脂的比较:模型组较正常组TC、TG、LDL-C水平明显升高;PCRarray结果显示:模型组与正常组相比,下调的≥4倍基因有17个,占总基因数的20%,如:ATP5A1、ATP5L、ATP5F1、ATP4A、ATP6V1E2等;上调≥4倍的基因转录占基因总数的10%,为OXA1L、NDUFB8、NDUFB9、NDUFS2、NDUFS8,涉及氢离子转运,琥珀酸脱氢酶亚基,NDUF脱氢酶亚基相关基因。结论:脾虚痰浊AS对心肌线粒体能量代谢的影响可能与涉及氢离子转运,琥珀酸脱氢酶亚基,NDUF脱氢酶亚基相关基因表达改变有关。

Objective： To observe mRNA expression genes related to Bama- mini pigs＇ heart mitochondrial energy metabolism signaling pathway in spleen deficiency and phlegm turbidity AS by PCRarray technology and explore the relationship between the expression of the gene of AS and spleen deficiency and phlegm turbidity. Methods： Ten male Bama- mini common grade pigs were randomly divided into control group,spleen deficiency and phlegm AS group. Spleen deficiency and phlegm AS group was given improper diet,overstrain combined with high fat diet. Automatic biochemical analyzer was used for determination of TC,TG,LDL- C. By Bama- mini pigs myocardial total RNA extraction and application of myocardial mitochondrial energy metabolism pathway PCR chip,determine spleen deficiency and phlegm syndrome AS related mRNA expression. Results： Comparison of Bama-mini pig＇s blood fat： spleen deficiency and phlegm syndrome AS patients compared with normal control group,triglycerides,total cholesterol and low density lipoprotein levels increased significantly. PCRarray results： model group＇s down- regulated larger than or equal to 4 times genes were 17,accounting for 20% of the total gene numbers,such as： ATP5a1,ATP5 L,ATP5F1,ATP4 A,ATP6V1E2. Those raised more than or equal to four times,accounting for 10% of the total number of genes,as OXA1 L,NDUFB8,NDUFB9,NDUFS2,NDUFS8,involving hydrogen ion transport and succinate dehydrogenase subunit,dehydrogenase,NDUF subunit related genes. Conclusion： The effect of hyperlipidemia on myocardial mitochondria may be possibly related to the changes of genes expressions which are related to the hydrogen ion transport,the succinate dehydrogenase subunit and the NDUF dehydrogenase subunit.